Fig. 2
From: LPA1-mediated inhibition of CXCR4 attenuates CXCL12-induced signaling and cell migration
LPA1 reduces CXCL12-induced cAMP suppression and ERK phosphorylation. A-D Inhibition of the CXCL12-induced cAMP response by CXCR4-LPA1 heteromers. A HEK293A cells were transfected with Myc-CXCR4 alone or together with increasing amounts of Flag-LPA1, and the effect of CXCL12 (30 nM) on forskolin (3 μM)-induced cAMP production was measured using a GloSensor cAMP assay. B HEK293A cells were transfected with Myc-CXCR4 (30 ng) alone or together with Flag-LPA1 (120 ng) and CXCL12-induced inhibition of cAMP production was measured. C HEK293A cells were transfected with Flag-LPA1 alone or together with increasing amounts of Myc-CXCR4, and the effect of alkyl-OMPT (1 μM) on forskolin (3 μM)-induced cAMP production was measured. D HEK293A cells were transfected with Flag-LPA1 (90 ng) alone or together with Myc-CXCR4 (120 ng) and inhibition of cAMP production by alkyl-OMPT was measured. Data represent the mean ± SEM of n = 3 to 5 independent experiments performed in triplicate. Statistical significance was tested using unpaired two-tailed Student’s t test (A, C) or two-way ANOVA followed by Bonferroni’s multiple comparison test (B, D). ***P < 0.001; ****P < 0.0001; ns, not significant. E–H Inhibition of CXCL12-induced ERK phosphorylation by CXCR4-LPA1 heteromers. HEK293A cells were transfected with CXCR4 (20 ng), LPA1 (80 ng), or both and ERK phosphorylation induced by CXCL12 (10 nM) (E) or alkyl-OMPT (1 μM) (F) was examined. (G) CXCL12 dose–response for ERK phosphorylation was measured in HEK293A cells expressing CXCR4 alone or together with LPA1. H Alkyl-OMPT dose–response for ERK phosphorylation was measured in HEK293A cells expressing LPA1 alone or together with CXCR4. Data represent the mean ± SD of n = 3 to 5 independent experiments. Statistical significance was tested using two-way ANOVA followed by Bonferroni’s multiple comparison test. *P < 0.05; ***P < 0.001; ns, not significant